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Signal amplification for thrombin impedimetric aptasensor : sandwich protocol and use of gold-streptavidin nanoparticles
Ocaña Tejada, Cristina (Universitat Autònoma de Barcelona. Grup Sensors i Biosensors)
Valle Zafra, Manuel del (Universitat Autònoma de Barcelona. Departament de Química)

Fecha: 2014
Resumen: In this work, we report a highly specific amplification strategy demonstrated for the ultrasensitive biosensing of thrombin with the use of gold-streptavidin nanoparticles (strep-AuNPs) and silver reduction enhancement. The biotinylated aptamer of thrombin was immobilized onto an avidin-graphite epoxy composite (AvGEC) electrode surface by affinity interaction between biotin and avidin; electrochemical impedance measurements were performed in a solution containing the redox marker ferrocyanide/ferricyanide. The change in interfacial charge transfer resistance (Rct) experimented by the redox marker, was recorded to confirm aptamer complex formation with target protein, thrombin (Thr), in a label-free first stage. A biotinylated second thrombin aptamer, with complementary recognition properties was then used in a sandwich approach. The addition of strep-AuNPs and silver enhancement treatment led to a further increment of Rct thus obtaining significant signal amplification. The AptThrBio1-Thr-AptThrBio2 sandwich formation was inspected by confocal microcopy after incubation with streptavidin quantum dots. In order to visualize the presence of gold nanoparticles, the same silver enhancement treatment was applied to electrodes already modified with the nanoparticle-sandwich conjugate, allowing direct observation by scanning electron microscopy (SEM). Results showed high sensitivity and selectivity for thrombin detection, with an improvement from ca. 4. 7 pM in a simple assay to 0. 3 pM in the amplified reported scheme.
Nota: Número d'acord de subvenció MICINN/CTQ2010-17099
Derechos: Tots els drets reservats.
Lengua: Anglès.
Documento: article ; recerca ; submittedVersion
Materia: Impedance ; Nanoparticles ; Aptamer ; Biosensor ; Thrombin ; Quantum dots
Publicado en: Biosensors and bioelectronics, Vol. 54 (2014) , p. 408-414, ISSN 1873-4235

DOI: 10.1016/j.bios.2013.10.068

21 p, 1.5 MB

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